Modeling Pulmonary HIV-Cytokine Interactions

肺部 HIV-细胞因子相互作用建模

• 批准号: 6800103
• 负责人: John E. Mittler
• 金额: $ 44.77万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6800103
• 关键词: AIDS therapy; HIV infections; alveolar macrophages; cell cell interaction; chemokine; clinical research; combination chemotherapy; cytokine; enzyme linked immunosorbent assay; human subject; latent virus infection; mathematical model; microarray technology; pathologic process; patient oriented research; respiratory infections; virus replication

DESCRIPTION (provided by applicant): Our proposal is devoted to defining the pathogenesis of HIV infection in the lung. It describes a series of in vivo and in vitro experiments oriented towards constructing mechanistic mathematical models to better understand cytokine networks and their role in the dynamics of HIV replication in the lung. Deciphering cytokine networks and their differential and combined effects on HIV-1 (HIV) replication in the lung presents a problem of enormous complexity. Our studies clearly show that pulmonary leukocytes can be a significant source of proinflammatory cytokines, chemokines and replicating virus in both children and adults, however, the actual impetus for lung involvement in HIV infection is not known. Our previous studies suggest that both viral and nonviral conditions that lead to a heightened state of macrophage activation favor the replication of HIV in the lung. This, along with alterations in concentration gradients and patterns of cytokine expression that promote cellular activation, may be crucial for establishing this niche. Our previous studies have also indicated that latently infected CD4vT cells in the blood contribute only a portion of the viral rebound following cessation of combination antiretroviral therapy (ART) and that other sites appear to be the source of much of the rebounding virus. Pulmonary macrophages, which are relatively long-lived and for which HIV infection is not overtly cytopathic, have been suggested as a possible persistent reservoir of HIV replication, including during long periods of ART. Hence, the more long-term objectives of our proposal are to better define the relationship between host and viral factors that promote HIV replication in the lung, particularly during ART. To do this, we will examine the range of HIV-specific host cells in the lungs of HIV-positive persons in relation to the stage of disease and cytokine/chemokine expression profiles. This will be accomplished using ELISA and cDNA microarray technologies and by in situ hybridization techniques that allow for the localization and quantification of specific viral and cytokine mRNAs within phenotypically-defined cells obtained from induced sputum and bronchoalveolar lavage (BAL). With these data we will construct and evaluate mechanistic and nonmechanistic mathematical models for the interaction between cells that replicate HIV and those that express specific chemokine and proinflammatory cytokines in the lung. We hypothesize that persons with the highest concentrations of HIV RNA in lung tissues or sputum/BAL fluids will have more vRNA+ alveolar macrophages in tissues and fluids and these cells will produce more virus on a per-cell basis that persons with lower viral load. We also hypothesize that in such persons the pulmonary microenvironment will be altered in favor of a Thl -type cytokine milieu. Last, we will test the hypothesis that the lung is a separate virologic compartment from blood by examining titers and burst size of replicating virus and viral decay kinetics during ART. As such, we propose a central role for alveolar macrophages in the persistence of HIV replication and subsequent pathogenesis. In particular, we hypothesize that infection persists in these cells in an active state under ART to which we will assess by examining viral sequence diversity and divergence in treated vs. untreated persons.

描述（由申请人提供）： 我们的建议致力于定义肺中HIV感染的发病机理。它描述了一系列旨在构建机械数学模型的体内和体外实验，以更好地了解细胞因子网络及其在肺中HIV复制动力学中的作用。解密的细胞因子网络及其对肺中HIV-1（HIV）复制的差异和综合作用提出了巨大的复杂性问题。我们的研究清楚地表明，肺白细胞可能是儿童和成人中促炎细胞因子，趋化因子和复制病毒的重要来源，但是，尚不清楚肺部参与HIV感染的实际动力。我们以前的研究表明，导致巨噬细胞活化状态升高的病毒和非病毒条件都有利于肺中HIV的复制。这以及促进细胞活化的细胞因子表达的浓度梯度和模式的改变可能对建立这种利基至关重要。我们先前的研究还表明，在停止抗逆转录病毒疗法（ART）后，血液中受感染的CD4VT细胞仅贡献了一部分病毒反弹，而其他部位似乎是许多反弹病毒的来源。肺巨噬细胞相对较长且艾滋病毒感染并非明显的细胞病变，被认为是可能的持续性HIV复制储层，包括在长期的艺术中。因此，我们提案的长期目标是更好地定义促进肺部艾滋病毒复制的宿主和病毒因素之间的关系，尤其是在艺术期间。为此，我们将检查HIV阳性人群中与疾病阶段和细胞因子/趋化因子表达谱相关的HIV特异性宿主细胞的范围。这将使用ELISA和cDNA微阵列技术以及原位杂交技术来实现，从而可以在从诱导的痰液和支气管腔液泡灌注（BAL）获得的表型定义的细胞中定位和定位和定量特定病毒和细胞因子mRNA。使用这些数据，我们将构建和评估复制HIV的细胞与表达肺中表达特定趋化因子和促炎性细胞因子的细胞之间相互作用的机械和非机械数学模型。我们假设肺组织中HIV RNA浓度最高的人或痰液/BAL液在组织和液体中将具有更多的VRNA+肺泡巨噬细胞，并且这些细胞会以较低病毒载荷的人为基础产生更多的病毒。我们还假设在这样的人中，肺微环境将改变，而有利于THL型细胞因子环境。最后，我们将通过检查滴度和ART期间复制病毒和病毒衰减动力学的滴度和爆发的大小来检验肺与血液中单独的病毒隔室的假设。因此，我们提出了肺泡巨噬细胞在HIV复制和随后的发病机理中的核心作用。特别是，我们假设这些细胞在ART下的活跃状态中持续存在感染，我们将通过检查病毒序列的多样性和在未经治疗的人与未经治疗的人中的差异来评估。